• Journal of Life Science
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• v.20 no.11
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• pp.1577-1581
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• 2010

Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.

• International Journal of Industrial Entomology and Biomaterials
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• v.30 no.1
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• pp.6-16
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• 2015

This study established the genetic characterisation of 10 microsporidian isolates infecting non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) collected from biogeographical forest locations in the State of Assam, India, using PCR-based markers assays: inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD). A Nosema type species (NIK-1s_mys) was used as control for comparison. The shape of mature microsporidian spores were observed oval to elongated, measuring 3.80 to $4.90{\mu}m$ in length and 2.60 to $3.05{\mu}m$ in width. Fourteen ISSR primers generated reproducible profiles and yielded 178 fragments, of which 175 were polymorphic (98%), while 16 RAPD primers generated reproducible profiles with 198 amplified fragments displaying 95% of polymorphism. Estimation of genetic distance coefficients based on dice coefficients method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was done to unravel the genetic diversity of microsporidians infecting Indian muga and eri silkworm. The similarity coefficients varied from 0.385 to 0.941 in ISSR and 0.083 to 0.938 in RAPD data. UPGMA analysis generated dendrograms with two microsporidian groups, which appear to be different from each other. Based on Euclidean distance matrix method, 2-dimensional distribution also revealed considerable variability among different identified microsporidians. Clustering of these microsporidian isolates was in accordance with their host and biogeographic origin. Both techniques represent a useful and efficient tool for taxonomical grouping as well as for phylogenetic classification of different microsporidians in general and genotyping of these pathogens in particular.

• Journal of Sericultural and Entomological Science
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• v.4
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• pp.97-101
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• 1965

This work was carried out to investigate the relation between the fat content and the metamorphosis as the silkworms grew. The results obtained are as follows: 1. The content, of which the dry matter had been 24% at the stage of newly hatched silkworms, were gradually decreased by feeding on, but suddenly increased to the 28.5% at the mature stage. Those decreased slightly from pupation to just before the moths, and again increased to the 32% of the dry matter when became the moths. 2. The content, of which the fat had been 16.2%, however, decreased as the silkworms grew. The fat content increased little by little from the first feeding to the vigorous eating, but the matured worm contained only 15.7% of the fat. 3. The fat content of both male and female showed an increase to 23.5% and 35.7%, respectively. After that, the contents of it decreased to 12% and suddenly increased to 26% just before the moth in female. In male the content of fat showed a slight increase till before the moth, and after moth became, it were 42.5%.

• Journal of Sericultural and Entomological Science
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• v.26 no.2
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• pp.1-6
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• 1984

The present study was made to establish the purification method of mature microsporidian spores by iso-density equilibrium technique using percoll and the serological discrimination by an indirect fluorescent antibody technique. The purification of new microsporidian spores took effect in the three steps purification method(precentrifugation-percoll iso-density equilibrium centrifugation-rising). There were clear differences in the size of spores between the new microsporidia and N. bombycis. The spores of N. bombycis is short elliptical of 2.07 in a ratio of length to width in diameter while that of new microsporidia is characterized with long elliptical shape which shows a ratio of 2.76 in length to width in diameter. The specific antigens of new microsporidia K79 spores was showed in the spores wall by the indirect fluorescent antibody reaction, and it was affected by the antibody against N. bombycis which antiserum was diluted in 1:20. It means that the new microsporidia K79 is serologically not identical to N. bombycis.

• Journal of Sericultural and Entomological Science
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• v.30 no.1
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• pp.8-19
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• 1988

The objective of the present study was to investigate the behavioral responses of sillkworm (Bombyx mori) larvae to: the dominant effect of walking distace: and the relationships between walking distance and the various economic characters on yellow sheets. The means of walking distances of progeny from F2 and F1 F1 and F2 generations, which was a hybrid between the long and the short walking distances, were shorter than those of parents. This may indicate that the genes for the short walking distances were partially dominant over those for the longwalking. Estimated broad sense heritability of larval walking distance was different for each larval instar, which fell into the range of 52∼84%. Genetic correlation was apparent between walking distances of all larval stages and some economic characters. The number of cocoons per liter and the period from incubation to emergence have shown the positive genetic correlation with walking distances. The cocoon yield, the sing cocoon weight, and the cocoon shell weight have shown to be in the negative correlation with walking distances. No correlation was observed between walking distances of the mature larvae and any economic characters. The analysis by the path coefficient showed negative direct effect of walking distances on the cocoon yield, and positive indirect effect on the single cocoon weight and the cocoon shell weight.

• Journal of Sericultural and Entomological Science
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• v.24 no.2
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• pp.55-72
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• 1983

These studies were done to find out any difference, ultrastructural, physical or chemical, between the shells of diapausing and non-diapausing eggs of the silkworm, Bombyx mori L. 1. From the electron-microscopic observation, the egg shells have four distinctive layers. In addition to the four layers, the shells in the diapausing eggs has another layer with low electron density on its surface. 2. The permeability of the egg shell to hydrochloride was much lower in diapausing egg than in non-diapausing egg. Also the permeability changed in the opposite directions with the egg age: the diapausing eggs decreased while non-diapausing ones increased. 3. The permeability increased when the diapausing egg shell was treated with HCl. When they were treated with ether, however, the increase in permeability was much smaller. It seems there was an ether soluble material involved in the content of the egg shell. 4. The diapausing eggs were also much more resistant to desiccation than the non-diapausing ones. The former, when treated with HCl or chilling, became less resistant to desiccation. 5. The positive histochemical response of the egg shell to PAS-Alcian blue and protein stainings suggests presence of abundant proteins and carbohydrates in the egg shell. On the other hand, the staining response to lipid was more positive in the inner layers than in the outer layer of the shell. 6. The egg shell adhesives seems to be mucopolysaccharides produced by colleterial glands, since the oviposited eggs showed a positive responses to carbohydrate and negative to lipid-staining chemicals, but not the mature oocytes in the ovarioles. 7. There were two bands on the electrophoretic pattern of the SH proteins extracted from the egg shells both in the diapausing egg and non-diapausing one: a slow moving major component and a fast moving minor one. However, the electrophoretic mobility showed a difference in the minor components between them. It is evident that the fast moving minor one of non-diapausing egg ran a little further than that of diapausing egg. 8. In amino acids analysis, no significant differences were found in their composition between diapausing and non-diapausing egg and SH proteins contain relatively more glycine and less cystine.